Accurately Managing Your Aseptic Processing By Using The Right Culture Media: A Study On 3P® SMART PLATES and BIOBALL® 

Introduction

Accuracy of Environmental Monitoring (EM) is key in maintaining control and preventing contamination issues in pharma companies, and culture media plays a pivotal role in Environmental Monitoring programs. 

Culture media must provide accurate, reliable, and consistent results. Their ability to promote microorganisms’ growth must be demonstrated and validated throughout the shelf-life of the culture media, using different batches, and intra- and inter-batch reproducibility tests inoculating strains described in the Pharmacopoeias and pharmaceutical plant environmental isolates are recommended to be carried out. 

This study demonstrates the excellent inter-batch reproducibility & specificity of 3P® SMART PLATES culture media – with over 750 batches tested within 18 months and tested with 19 Pharma plant environmental isolates.

Materials And Methods

Growth Promotion Testing was performed using

• TSA 3P ref. 43169 
• TSA 3P with Neutralizers ref. 43819 
• COUNT-TACT® 3P® ref. 43699 

750 batches were analyzed (250 for each reference) and produced over 18 months. Each batch was tested twice. 

The Recovery Rate (RR) was calculated compared to a previous validated batch of the same reference. 

To minimize variability associated with the microorganisms' growth in broth culture, BIOBALL® freeze-dried microorganisms were used. Each lyophilized BIOBALL® contains a quantified number of microorganisms and provides a quick and ready-to-use solution, specifically designed for Growth Promotion Testing efficiency. BIOBALL® is an ISO:17034 accredited “Certified Reference Material” with strains described in pharmacopoeias harmonized chapters USP <61>, EP 2.6.12, and JP 4.05 (Table 1).

Table 1: Summary of BIOBALL® strains used for the study, and incubation conditions.

Performance Summary

Table 2: Summary of average recovery rates of 250 batches of each culture medium reference, for E. coli and pharmacopoeia harmonized chapters strains.

The study performed with TSA 3P, TSA3P with Neutralizers, and COUNT-TACT® 3P® on 6 pharmacopoeia strains showed that the mean recovery rates for the 250 batches analyzed over 18 months were close to 100%, demonstrating that these media have a strong reproducibility between the batches tested and over time.  

The FDA has issued warning letters about failure to use environmental isolates in microbiological testing. Other regulatory bodies increasingly request the same. Environmental isolates are the best way to challenge culture media and validation studies. Based on US FDA guidance, Quality Control laboratories are expected to determine if compendial organisms sufficiently represent production-related isolates and suggest adding environmental isolates to the growth promotion challenge if needed. 

Materials And Methods

19 environmental isolates from different parts of the world (Table 3), in BIOBALL® ready-to-use format, were tested on:

• TSA3P ref. 43169 
• TSA3P with Neutralizers ref. 43819 
• COUNT-TACT® 3P® ref. 43699 

For each culture media, three different batches were tested and performed in triplicate. The Recovery Rates were calculated comparing the mean counts obtained on their reference growth media selected for their ability to recover these stressed wild strains.

Table 3: Summary table of environmental isolates.

All isolates grew well as indicated by the high Recovery Rates calculated compared to the counts on their reference growth media (illustrated in Figures 4 and 5), demonstrating:

• That the growth performance for 3P® culture media is similar to the media that best enhances the growth of environmental isolates from pharmaceutical plants. 
• The excellent growth performance of 3P® culture media for environmental isolates. These media can detect a wide range of microorganisms present worldwide; therefore ensuring that potential contamination in an aseptic area might be detected when using 3P® culture media.

Figure 4 and 5: Growth performance (Recovery Rate in %) of environmental isolates according to microbial-group and geographical location.

This study demonstrates the excellent and robust performances of 3P® culture media in terms of reproducibility and specificity using both standard Pharmacopoeia strains and environmental isolates.  

These results support that the risk of false negative results is dramatically reduced, bringing confidence to pharmaceutical companies and helping ensure patient safety.  

3P® Culture media are designed for use by pharmaceutical manufacturers in their most critical manufacturing environments and are optimized for maximum specificity and reproducibility within the same lot and across multiple batches over time. Flexible storage conditions from 2-25°C and sample security achieved by locking plates enable 3P® culture media to be an effective component for risk-based environmental monitoring programs. 

The ATCC trademark and trade name and any and all ATCC catalog numbers are trademarks of the American Type Culture Collection. 

NCTC  and NCPF are a registered trademarks of the UK Health Security Agency.